The objective of this work was to determine, in laboratory, the displacement index and the separation index of earthworms (Eudrilus eugeniae) from the humus, to define the characteristics of the electric pulses that allow better efficiency in the separation of earthworms in pilot scale. The tests presented the following characteristics: (i) tension - 100 to 500 V; (ii) pulse level - 2.3 and 4 ms; (iii) frequency - 1 and 5 Hz; (iv) exhibition time - 1, 2, 3 and 4 h. It was verified that the earthworms submitted to the electric pulses preferentially move towards the cathode. For the studied situations the highest value obtained for the displacement index was 78.18%, while the separation index was 63.37%. The utilization of controlled electric pulses for the earthworm separation from the substrate generate mean gain along the estimated useful life of, around 26% of economic savings.